文章摘要

丙型肝炎病毒蛋白对肝细胞RASSF2 mRNA表达的影响及其机制

作者: 1陈炜, 1冯德云, 1李波, 1王颖
1 中南大学湘雅医院 病理科，湖南 长沙 410008
通讯: 王颖 Email: 20527296@qq.com
DOI: 10.3978/.10.3978/j.issn.1005-6947.2017.05.010
基金: 湖南省自然科学基金资助项目, 14JJ3048

摘要

目的：探讨丙型肝炎病毒（HCV）蛋白NS3、Core、NS5A对肝细胞RASSF2 mRNA表达及RASSF2A启动子甲基化的状态的影响。方法：用RT-PCR检测分别转染NS3、Core、NS5A表达质粒的肝QSG7701细胞（NS3/QSG7701、Core/QSG7701、NS5A/QSG7701）以及正常肝细胞L02中RASSF2 mRNA的表达；用甲基化特异性PCR检测NS3/QSG7701、Core/QSG7701及NS5A/QSG7701细胞中RASSF2A启动子甲基化的状态，以及去甲基化药物5-aza-dC处理后，各细胞RASSF2 mRNA表达情况与生物学行为的变化。结果：与正常肝细胞L02比较，NS3/QSG7701、Core/QSG7701、NS5A/QSG7701细胞中RASSF2 mRNA的表达均明显降低（均P<0.05）；3种细胞的RASSF2A启动子均发生完全甲基化。经5-aza-dC处理后，RASSF2 mRNA的表达在NS3/QSG7701和Core/QSG7701细胞中明显上调（均P<0.05），但在NS5A/QSG7701细胞中无明显变化（P>0.05）；NS3/QSG7701和Core/QSG770细胞经5-aza-dC处理后，增殖率下降、凋亡率增加（均P<0.05）。结论：NS3、Core、NS5A能通过RASSF2A启动子甲基化，且NS5A还通过其他机制降低RASSF2表达，该作用可能参与了HCV相关肝细胞癌的发生。
关键词： 肝肿瘤 病毒非结构蛋白质类 甲基化 肿瘤抑制蛋白质类

Influences of hepatitis C virus proteins on RASSF2 mRNA expression in hepatic cells and their mechanisms

Authors: 1CHEN Wei, 1FENG Deyun, 1LI Bo, 1WANG Ying
1 Department of Pathology, Xiangya Hospital, Central South University, Changsha 410008, China

CorrespondingAuthor:WANG Ying Email: 20527296@qq.com

Abstract

Objective: To investigate the influences of hepatitis C virus (HCV) proteins NS3, Core and NS5A on RASSF2 mRNA expression and methylation status of RASSF2A promoter in hepatic cells. Methods: The RASSF2 mRNA expressions in hepatic cells respectively transfected with NS3-, Core- and NS5A-expression plasmids (NS3/QSG7701, Core/QSG7701 and NS5A/QSG7701) as well as normal hepatic cell line L02 were determined by RT-PCR method. The methylation statuses of RASSF2A promoter in NS3/QSG7701, Core/QSG7701 and NS5A/QSG7701 cells were examined by methylation-specific PCR, and the changes in their RASSF2 mRNA expressions and biological behaviors after treatment with demethylation agent 5-aza-dC were observed. Results: The RASSF2 mRNA expressions were significantly decreased in all NS3/QSG7701, Core/QSG7701 and NS5A/QSG7701 cells compared with L02 cells (all P<0.05), and the RASSF2A promoters were fully methylated in all the three types of cells. After 5-aza-dC treatment, the RASSF2 mRNA expression was upregulated in NS3/QSG7701 and Core/QSG7701 cells (both P<0.05), but did not change in NS5A/QSG7701 cells (P>0.05); the proliferation rate was decreased and apoptosis rate was increased in NS3/QSG7701 and Core/QSG7701 cells5-aza-dC treatment (both P<0.05). Conclusion: NS3, Core and NS5A can reduce RASSF2 expression through RASSF2A promoter methylation and also through other mechanism by NS5A. This effect may probably contribute to the carcinogenesis of HCV-associated hepatocellular carcinoma.
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